Comparison of scanning Kelvin probe with SEM/EPMA techniques for fingermark recovery from metallic surfaces.

Most traditional techniques to recover latent fingermarks from metallic surfaces do not consider the metal surface properties and instead focus on the fingermark chemistry. The scanning Kelvin probe (SKP) technique is a non-contact, non-destructive method, used under ambient conditions, which can be utilised to recover latent prints from metallic surfaces and does not require any enhancement techniques or prevent subsequent forensic analysis. Where a fingermark ridge contacted the metal, the contact potential difference (CPD) contrast between the background surface and the fingermark contact area was 10-50mV. Measurements were performed on the untreated brass, nickel-coated brass and copper metal surfaces and compared to traditional forensic enhancement techniques such as Vacuum Metal Deposition (VMD) using Au-Zn and Au-Ag. Using VMD, the CPD change ranged from 0 to 150mV between the dissimilar metal surfaces affected by the fingermark. In general, SKP worked best without additional enhancement techniques. Scanning Electron Microscope (SEM) scans were used to identify the fingermark contact areas through a sodium, chlorine and oxygen electron probe micro-analyzer (EPMA). The fingermark was observed in the backscattered electron image as the carbon deposits scattered the electrons less than the surrounding metal surface. The fingermark is shown clearly in a Cathodoluminescence scan on the copper sample as it blocks the photon emission at band gap (2.17eV) from the underlying copper oxide (Cu2O) surface. For the first time, SEM, EPMA and Cathodoluminescence techniques were compared to SKP data. Visible and latent fingermarks were tested with latent, eccrinous fingermarks more easily imaged by SKP. Results obtained were very encouraging and suggest that the scanning Kelvin probe technique, which does not need vacuum, could have a place as a first stage analysis tool in serious crime investigation.

Long term follow up of men with Alfuzosin who voided successfully following acute urinary retention*.

INTRODUCTION: Acute urinary retention (AUR) is one of the commonest causes of admission in urology ward and successful voiding with alpha-blockers has been reported. However, long-term efficacy of Alfuzosin, following an episode of AUR is lacking. This is a continuation of our earlier reported study. We report the results of a 4 year follow-up on patients who were on Alfuzosin SR 5 mg BD, following resumption of voiding after an episode of AUR. PATIENTS AND METHODS: A total of 33 patients voided successfully following AUR in our original study. These patients continued on Alfuzosin SR 5 mg BD and were assessed at 2 and 4 years. Symptomatic assessment was performed with IPSS and QOL symptom score and objective assessment was with urinary flow rate and post void residual volume. Patients who continued to deteriorate symptomatically and objectively or developed further AUR were listed for surgery. RESULTS: Out of 33 patients, 28 patients were followed up at 2 years (three patients died due to various medical reasons and two did not attend for follow-up). Nineteen patients (68%) underwent transurethral resection of prostate (TURP) for severe lower urinary tract symptom (LUTS) . The mean peak flow rate at 2 years was 8.4 ml/s and the mean residual volume was 112 ml. Ten patients attended for follow up at 4 years. The mean flow rate was 5.17 ml/s and the mean post-void residual volume was 101 ml. Four patients underwent TURP for severe outflow symptoms. At 4 years follow up 24 out of 30 patients (80%) on Alfuzosin needed TURP. CONCLUSIONS: These data do not support the long term use of alpha-blockers in patients who voided successfully after acute urinary retention.

NAD(P)H:Quinone oxidoreductase-1 C609T polymorphism analysis in human superficial bladder cancers: relationship of genotype status to NQO1 phenotype and clinical response to Mitomycin C.

NAD(P)H:Quinone oxidoreductase-1 (NQO1) has been implicated in the bioreductive activation of the clinically active anticancer drug Mitomycin C (MMC) and a polymorphic variant of NQO1 which lacks functional enzyme activity (NQO1*2) has been linked with poor survival in patients treated with MMC. The relationship between NQO1 activity and cellular response to MMC is however controversial and the aim of this study was to determine whether the response of bladder cancer patients to MMC can be forecast on the basis of NQO1*2 genotype status. Genomic DNA was extracted from formalin-fixed, paraffin-embedded tissue from 148 patients with low to intermediate grade (G1/G2) superficial (Ta/T1) bladder cancers and NQO1*2 genotype status determined by PCR-RFLP. NQO1*2 genotype status was retrospectively compared with clinical response to intravesical administered MMC with the primary end-point being time to first recurrence. NQO1 phenotype was determined by immunohistochemistry. Of the 148 patients genotyped, 85 (57.4%) were NQO1*1 (wild-type), 59 (39.8%) were NQO1*1/*2 (heterozygotes) and 4 (2.7%) were NQO1*2/*2. No NQO1 protein expression was detected in NQO1*2/*2 tumours. A broad spectrum of NQO1 protein expression existed in tumours genotyped as NQO1*1 and NQO1*1/*2 although tumours with NQO1*1 typically expressed higher NQO1 protein. A poor correlation existed between NQO1*2 genotype status and clinical response to MMC. The results of this retrospective study suggest that tailoring MMC therapy to individual patients with superficial bladder cancer on the basis of NQO1 genotype status is unlikely to be of clinical benefit.

Immunohistochemical analysis of NAD(P)H:quinone oxidoreductase and NADPH cytochrome P450 reductase in human superficial bladder tumours: relationship between tumour enzymology and clinical outcome following intravesical mitomycin C therapy.

A central theme within the concept of enzyme-directed bioreductive drug development is the potential to predict tumour response based on the profiling of enzymes involved in the bioreductive activation process. Mitomycin C (MMC) is the prototypical bioreductive drug that is reduced to active intermediates by several reductases including NAD(P)H:quinone oxidoreductase (NQO1) and NADPH cytochrome P450 reductase (P450R). The purpose of our study was to determine whether NQO1 and P450R protein expression in a panel of low-grade, human superficial bladder tumours correlates with clinical response to MMC. A retrospective clinical study was conducted in which the response to MMC of 92 bladder cancer patients was compared to the immunohistochemical expression of NQO1 and P450R protein in archived paraffin-embedded bladder tumour specimens. A broad spectrum of NQO1 protein levels exists in bladder tumours between individual patients, ranging from intense to no immunohistochemical staining. In contrast, levels of P450R were similar with most tumours having moderate to high levels. All patients were chemotherapy naïve prior to receiving MMC and clinical response was defined as the time to first recurrence. A poor correlation exists between clinical response and NQO1, P450R or the expression patterns of various combinations of the 2 proteins. The results of our study demonstrate that the clinical response of superficial bladder cancers to MMC cannot be predicted on the basis of NQO1 and/or P450R protein expression and suggest that other factors (other reductases or post DNA damage events) have a significant bearing on tumour response.

Detection of (NAD(P)H:Quinone oxidoreductase-1, EC 1.6.99.2) 609C-->T and 465C-->T polymorphisms in formalin-fixed, paraffin-embedded human tumour tissue using PCR-RFLP.

NQO1 is a cytosolic flavoprotein that plays a dual role in the detoxification of potentially carcinogenic compounds and the bioreductive activation of quinone based anticancer drugs. Two polymorphic variants of NQO1 exist (NQO1*2 and NQO1*3) which cause significant phenotypic reductions in NQO1 protein content and activity. Current methods for detecting NQO1 polymorphisms commonly use PCR-RFLP techniques and have exclusively used DNA isolated from fresh tissues. This study describes a method that is suitable for analysing NQO1 polymorphisms in genomic DNA isolated from formalin-fixed paraffin-embedded tissue. The method utilises two rounds of PCR amplification using a nested primer strategy that generates specific PCR products followed by RFLP analysis using either Hinf1 (for NQO1*2) or Msp1 (for NQO1*3). Whilst existing methods proved unsatisfactory (low product yield and poor specificity), the nested primer strategy produced good quality PCR products suitable for RFLP analysis and genotyping of NQO1*2 and NQO1*3 in archival tissue samples. The ability to utilise the vast archives of human tissue held by pathology laboratories would be of considerable benefit as retrospective studies comparing NQO1 genotype status, patient history and treatment outcomes could be conducted.

Randomised, placebo controlled, double blind study of alfuzosin SR in patients undergoing trial without catheter following acute urinary retention.

INTRODUCTION: Acute urinary retention caused by bladder outlet obstruction resulting from prostatic enlargement is one of the commonest causes for acute admission to urology wards. More recently, there has been a trend to commence treatment with alpha-blockers after catheterisation followed by a trial without catheter (TWOC), in the hope that surgery may be avoided in a significant proportion of patients. There is no conclusive evidence of the efficacy of this treatment. We conducted a study to evaluate the efficacy of using the alpha-blocker alfuzosin SR in patients with acute urinary retention. PATIENTS AND METHODS: All patients presenting with acute urinary retention to our unit were included in the trial. Exclusion criteria included patients with known bladder or prostate malignancy, bladder calculi, urinary tract infections, urethral stricture or patients on alpha-blockers. A total of 81 patients consented and were randomised. Sixty-two patients completed the study. The retention volume was recorded. Trial medicine was recorded on a twice-daily dose and the first TWOC was carried out after a minimum of three doses or 36 hours after admission. TWOC was considered successful on voiding with a residual volume of <200 ml. Unsuccessful patients were recatheterised and discharged home on trial medication, and called for a second TWOC after 2 weeks. Successful patients were continued on alpha-blockers and failures were put on the operating list for TURP. Patients on active treatments were reviewed at 2 year. RESULTS: Of the 34 patients treated with alfuzosin SR, 17 (50%) resumed voiding and of the 28 patients from placebo group, 16 (57%) voided successfully. All 33 patients were continued open labelled on alfuzosin SR 5mg BD. Out of 33 patients, 13 (43%) had TURP within first year after TWOC and three died due to various medical causes. Out of remaining 17 patients, 15 attended for follow-up. The mean peak flow rate was 8.4 ml/s and the mean residual volume was 112 ml. Six patients (40%) required TURP for severe lower urinary tract symptoms (LUTS). So out of 28 patients followed at 2 year, 19 (68%) had TURP. CONCLUSIONS: These data do not support the routine use of alpha-blockers in patients with acute urinary retention. Also continuing use of alpha-blockers does not seem to prevent further requirements of TURP, although larger studies are needed to support this.

A novel strategy for NQO1 (NAD(P)H:quinone oxidoreductase, EC 1.6.99.2) mediated therapy of bladder cancer based on the pharmacological properties of EO9.

The indolequinone EO9 demonstrated good preclinical activity but failed to show clinical efficacy against a range of tumours following intravenous drug administration. A significant factor in EO9's failure in the clinic has been attributed to its rapid pharmacokinetic elimination resulting in poor drug delivery to tumours. Intravesical administration of EO9 would circumvent the problem of drug delivery to tumours and the principal objective of this study is to determine whether or not bladder tumours have elevated levels of the enzyme NQO1 (NAD(P)H:quinone oxidoreductase) which plays a key role in activating EO9 under aerobic conditions. Elevated NQO1 levels in human bladder tumour tissue exist in a subset of patients as measured by both immunohistochemical and enzymatic assays. In a panel of human tumour cell lines, EO9 is selectively toxic towards NQO1 rich cell lines under aerobic conditions and potency can be enhanced by reducing extracellular pH. These studies suggest that a subset of bladder cancer patients exist whose tumours possess the appropriate biochemical machinery required to activate EO9. Administration of EO9 in an acidic vehicle could be employed to reduce possible systemic toxicity as any drug absorbed into the blood stream would become relatively inactive due to an increase in pH.

Risk factors and behaviors associated with separation anxiety in dogs.

OBJECTIVES: To determine potential risk factors and behaviors associated with separation anxiety and develop a practical index to help in the diagnosis of separation anxiety in dogs. DESIGN: Case-control study. ANIMALS: 200 dogs with separation anxiety and 200 control dogs with other behavior problems. PROCEDURES: Medical records were reviewed for signalment, history of behavior problems, home environment, management, potentially associated behaviors, and concurrent problems. RESULTS: Dogs from a home with a single adult human were approximately 2.5 times as likely to have separation anxiety as dogs from multiple owner homes, and sexually intact dogs were a third as likely to have separation anxiety as neutered dogs. Several factors associated with hyperattachment to the owner were significantly associated with separation anxiety. Spoiling activities, sex of the dog, and the presence of other pets in the home were not associated with separation anxiety. CONCLUSIONS AND CLINICAL RELEVANCE: Results do not support the theory that early separation from the dam leads to future development of separation anxiety. Hyperattachment to the owner was significantly associated with separation anxiety; extreme following of the owner, departure cue anxiety, and excessive greeting may help clinicians distinguish between canine separation anxiety and other separation-related problems.

Telomerase activity in malignant and benign bladder conditions.

This study investigated whether the enzyme telomerase is active in bladder tumours, whether there is any correlation between activity and grade, and whether the enzyme is expressed in non-malignant conditions. Fifty-two patients undergoing cystoscopy or TURBT at a district general hospital were included, 25 with current bladder tumours, 13 with previous but no current tumours, and 14 with non-malignant pathology. Specimens were analysed by the telomerase repeat amplification protocol (TRAP assay), a highly-sensitive polymerase chain reaction (PCR)-based assay, and a commercially-available ELISA kit. Telomerase activity was detected in 80% of bladder tumours, more frequently in moderate- or poorly-differentiated (93%) than well-differentiated (56%) tumours. Activity was not uniform across individual tumours. Telomerase was also frequently (61%) detected in inflammatory lesions found in patients being followed up for previous bladder tumours, and in two (14%) patients with benign pathologies. In conclusion, telomerase was frequently but not uniformly detected in bladder tumours; its presence was not specific to malignancy. There is a possible correlation between tumour grade and telomerase activity.

Role of P-glycoprotein in chemoresistant superficial bladder tumours.

OBJECTIVE: The drug resistance at cellular level is mediated by P-glycoprotein (P-G), which is variably expressed in bladder tumours. The effect of intravesical chemotherapy on P-G status was studied in chemoresistant and recurrent superficial tumours which progressed to metastasis or needed further treatment. METHODS: Archival histological materials of 14 patients who received intravesical epirubicin for recurrent superficial transitional cell carcinoma of the bladder were studied for the presence of P-G using monoclonal antibody JSB-1. RESULTS: Four patients showed complete absence of P-G following chemotherapy although only 2 patients were recurrence-free. In 4 patients with extravesical metastasis, there was no evidence of increased P-G expression. CONCLUSIONS: The P-glycoprotein expression is not related to histological grading or clinical progression of bladder tumours.

Ultrasound in urological emergency: results of self audit and implications for training.

The efficacy of ultrasound examination by the trainee urologists in the management of urological emergencies admitted in a district general hospital was studied. Fifty patients (100 kidney units) had renal ultrasound performed by urological trainees on acute admission. The results were compared with subsequent definitive radiological investigations. On analysis of 100 renal units there were 7 discordant results, 2 false negatives and 5 false positives achieving 97% specificity (95% confidence interval 93% to 100%) and 84% sensitivity (95% confidence interval 71% to 97%). Adopting 50-patient analysis there was 89% specificity (95% confidence interval 74% to 100%) and 84% sensitivity (95% confidence interval 71% to 97%). These intervals indicate the levels of success to be expected in future studies. The study shows that urological trainees can use ultrasound with high levels of accuracy improving patient management.

Radiographer-performed abdominal and pelvic ultrasound: its value in a urology out-patient clinic.

OBJECTIVE: To assess the efficacy of radiographer-performed ultrasound examination as a routine investigative procedure in a urological out-patient clinic. PATIENTS AND METHODS: A total of 151 patients attending a District General Hospital Urological Out-patient Department underwent an ultrasound examination in the clinic. RESULTS: Diagnosis by ultrasound was achieved in 93% of patients. The remaining patients underwent further investigations. Two (1%) patients with normal scans had small bladder tumours. Subsequent intravenous urography in these individuals showed normal upper tracts. CONCLUSION: Abdominal and pelvic ultrasound examination performed in the urological out-patient clinic on unprepared patients was the only investigation necessary for evaluation of common problems such as non-specific urinary symptoms, recurrent urinary tract infections and bladder outlet obstruction.

Growth characteristics of early passage cell lines compared with established TCC bladder lines.

The growth patterns of established cell lines from bladder transitional cell carcinoma (TCC) were compared with early passage cell lines. The growth of established cell line 5637 was uninhibited in both serum free (basal) and serum containing media. The early passage line (DR) grew only in serum containing medium. This confirms the unreliability of results from biological studies on established (continuous) cell lines.

Echocardiographic evidence for myocardial failure induced by taurine deficiency in domestic cats.

Dietary taurine-deficiency is a cause of dilated cardiomyopathy (DCM) in cats. While the incidence of clinical cases of feline DCM has markedly decreased since the association between DCM and taurine-deficiency was first recognized, not all cats maintained on taurine-deficient diets develop DCM. The objective was to temporally evaluate left ventricular (LV) function using M-mode echocardiography in 23 cats maintained on a taurine-deficient diet; 20 time-matched, taurine-supplemented cats served as controls. The duration of feeding trials ranged from 6-15 months. No diminution of myocardial function was recorded in a small number of taurine-deficient cats whereas cardiac performance in some taurine-deficient cats diminished to levels characteristic of DCM. Of the taurine-deficient cats, 17 (74%) experienced a greater than 25% reduction in fractional shortening and 21 (91%) had a greater than 25% increase in LV end-systolic short-axis diameter. On average, LV end-systolic short-axis diameter increased by 70% and fractional shortening decreased by 37% in taurine-deficient cats. Mean velocity of circumferential fiber shortening was similarly reduced in taurine-deficient cats. The greatest rate of change in M-mode echocardiographic variables occurred during the first four months on the taurine-deficient diet. Dietary taurine deficiency leads to a spectrum of changes in myocardial function in domestic cats. While DCM is observed in some cats, decreased systolic pump function and increased LV end-systolic short-axis diameter are more consistent findings.